Novel peptide and use thereof
一种组合物、药学的技术,应用在治疗和/或预防软骨损伤或关节炎的新型肽领域,能够解决不容易获得足够数量的自体细胞等问题
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Embodiment 1
[0071] [Example 1] Preparation of peptide
[0072] At the request of the present inventors, a peptide having the amino acid sequence shown in SEQ ID NO. 1 (Glu-Leu-His-Leu-Asp) was prepared by Peptron Corporation (South Korea). Specifically, using an automated peptide synthesizer (ASP48S, Peptron Inc.), the amino acid units were linked one by one from the C-terminus by Fmoc SPPS (9-fluorenylmethoxycarbonyl solid-phase peptide synthesis method).
[0073] Use NH 2-His(Trt)-2-chloro-trityl resin, the first amino acid at the C-terminus of the peptide was attached to the resin. All amino acids used to synthesize peptides are protected by trityl (Trt), tert-butoxycarbonyl (Boc), tert-butyl (t-Bu), etc., where the N-terminus is protected by Fmoc, and all amino acids are removed in acid Residues. Using 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) / hydroxybenzotriazole (HOBT) / N-methylmorphine phenoline (NMM) as the coupling reagent. (1) Dissolve th...
Embodiment 2-12
[0074] [Example 2-12] Preparation of Peptides
[0075] The peptides of Examples 2-12 were prepared in the same manner as in Example 1, except that the amino acid sequences listed in Table 1 below were used instead of the amino acid sequences in Example 1.
[0076] Table 1
[0077] Example serial number
Embodiment 13
[0078] [Example 13] Confirmation of cartilage regeneration effect with damaged cartilage tissue explants
[0079] Articular cartilage explants were prepared from hoof joints of cows younger than 3 years old within one hour of slaughter at a regional slaughterhouse (located in Ojeong-dong, Daejeon, South Korea). For this purpose, the cartilage part other than the bone was evenly excised from the joint using a scalpel, and cut into a size of about 3 mm x 3 mm to prepare cartilage tissue explants. Using a syringe (21G) with a truncated and ground tip, the lesion area was prepared by uniformly making vertical holes in the surface of the center of the explant.
[0080] The perforated tissue explants were divided into 4 groups including the control group. Each group was placed in Dulbecco's modified Eagle's medium / F12 (DMEM / F12, 1:1, Welgene) supplemented with ascorbate (50 μg / ml, Sigma) and 10% fetal bovine serum (FBS, Invitrogen). ); then treated with 25 μM of the peptide of Exa...
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